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Comparative analysis of the survival and regeneration potential of juvenile and matured earthworm, Eudrilus eugeniae, upon in vivo and in vitro maintenance

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Abstract

Eudrilus eugeniae is a clitellum-dependent earthworm that requires intact clitellum segments for its survival and regeneration. The present study aims to interconnect the survival and regeneration ability that varies between in vivo and in vitro maintenance upon different sites of amputation. The amputated portion of the worm that possesses intact clitellum (13th–18th segments) survived and had the potential to regenerate, whereas worms with partial or without clitellum segments only survived and were unable to regenerate. Besides segment length and clitellum segments, clitellum factors also determined the survival, blastemal initiation and differentiation potential. The survivability and regeneration potential of worms were augmented upon in vitro maintenance. Notably, the amputated segments (1st–10th segments) and posterior segments of similar length, which usually die within the 4th day in vivo, survived for more than 60 days in vitro but lacked the regeneration ability. On the other hand, the amputated posterior segments (30th to 37th segments) from juvenile worms, maintained in in vitro condition, survived and initiated blastema with multiple buds but lacked the ability to regenerate. Interestingly, the equal half of adult worm blastema that is maintained in in vitro conditions were able to form the blastema-like structure with the help of a unique stick. The anterior blastema failed to retain the regenerative structure but the posterior portion of the amputated blastema, which is also associated with a small portion of the body segment, showed the ability to retain the regenerative structure. Our results conclude that the survivability is enhanced upon in vitro maintenance and this condition favours the adult dedifferentiated blastemal and stem cell–enriched juvenile posterior segments to form a regenerative blastema.

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All data generated or analysed during this study are included in this published article.

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Acknowledgements

The authors thank ‘International Research Centre (IRC) of Sathyabama Institute of Science and Technology, Chennai’ for  providing support to carry out the research work.

Funding

The ‘Department of Science and Technology—Science and Engineering Research Board (DST-SERB, Ref. No. ECR/2016/000956), New Delhi, India’.

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Authors and Affiliations

  1. Regeneration and Stem Cell Biology Lab, Centre for Molecular and Nanomedical Sciences, International Research Centre, Sathyabama Institute of Science and Technology (Deemed to Be University), Chennai, Tamil Nadu, 621306, India

    Kamarajan Rajagopalan, Jackson Durairaj Selvan Christyraj, Karthikeyan Subbiahanadar Chelladurai & Johnson Retnaraj Samuel Selvan Christyraj

  2. Department of Biotechnology, Vel Tech Rangarajan Dr. Sagunthala R&D Institute of Science and Technology, Avadi, Chennai, Tamil Nadu, India

    Jemima Kamalapriya John Samuel Gnanaraja

  3. School of Health Sciences, Purdue University, West Lafayette, USA

    Karthikeyan Subbiahanadar Chelladurai

Authors
  1. Kamarajan Rajagopalan
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  2. Jackson Durairaj Selvan Christyraj
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  3. Karthikeyan Subbiahanadar Chelladurai
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  4. Jemima Kamalapriya John Samuel Gnanaraja
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  5. Johnson Retnaraj Samuel Selvan Christyraj
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Contributions

Mr. Kamarajan Rajagopalan contributed to the writing of the original draft, project conceptualization, data collection and the development of figures and tables; Dr. Jackson Durairaj Selvan Christyraj was involved in writing the original draft, project conceptualization, performance, supervision and administration; Karthikeyan Subbiahanadar Chelladurai and Jemima Kamalapriya John Samuel Gnanaraja were involved in reviewing the results and language editing. Johnson Retnaraj Samuel Selvan Christyraj was involved in writing the original draft and project conceptualization.

Corresponding authors

Correspondence to Jackson Durairaj Selvan Christyraj or Johnson Retnaraj Samuel Selvan Christyraj.

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Ethics approval

The experiments are carried out using lower invertebrate earthworm; therefore, ethical statement is not needed. Necessary care is taken in experimental procedure that is intended to avoid unnecessary pain and suffering to the experimental animals.

Conflict of interest

The authors declare no competing interests.

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Rajagopalan, K., Christyraj, J.D.S., Chelladurai, K.S. et al. Comparative analysis of the survival and regeneration potential of juvenile and matured earthworm, Eudrilus eugeniae, upon in vivo and in vitro maintenance. In Vitro Cell.Dev.Biol.-Animal 58, 587–598 (2022). https://doi.org/10.1007/s11626-022-00706-6

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